NDLI: A haptotaxis assay for leukocytes based on surface-bound chemokine gradients.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Rink, Jan Rink, Ina Schmitz, Katja Helmer, Dorothea Sachs, Daniel
Description	Author Affiliation: Rink I ( Technische Universität Darmstadt, Clemens-Schöpf-Institut für Organische Chemie und Biochemie, 64287 Darmstadt, Germany); Rink J ( Technische Universität Darmstadt, Ernst-Berl-Institut für Technische und Makromolekulare Chemie, 64287 Darmstadt, Germany); Helmer D ( Karlsruhe Institute of Technology, Institute of Microstructure Technology, 76344 Eggenstein-Leopoldshafen, Germany.); Sachs D ( Technische Universität Darmstadt, Clemens-Schöpf-Institut für Organische Chemie und Biochemie, 64287 Darmstadt, Germany); Schmitz K ( Technische Universität Darmstadt, Clemens-Schöpf-Institut für Organische Chemie und Biochemie, 64287 Darmstadt, Germany)
Abstract	The migration of leukocytes in response to chemokine gradients is an important process in the homeostasis of the human immune system and inflammation. In vivo the migration takes place on the surface of the endothelium to which the chemokine gradient is immobilized via interaction with glycosaminoglycans. To study leukocyte migration in response to surface-bound chemokines, we generated chemokine gradients by a simple stamping method: agarose stamps were soaked with chemokine solution to form continuous chemokine gradients by diffusion. These gradients could be easily transferred to a petri dish surface by stamping. We show that neutrophil granulocytes recognize these gradients and migrate toward increasing chemokine concentrations dependent on the slope of the gradient. Single-cell responses were recorded, and statistical analyses of cell behavior and migration were performed. For analysis of chemotaxis/haptotaxis, we propose a chemotactic precision index that is broadly applicable, valid, and allows for a straightforward and rapid quantification of the precision by which cells follow the direction of a given gradient. The presented technique is very simple, cost-efficient, and can be broadly applied for generating defined and reproducible immobilized gradients of almost any protein on surfaces, and it is a valuable tool to study haptotaxis.
ISSN	00221767
e-ISSN	15506606
Journal	The Journal of Immunology
Issue Number	11
Volume Number	194
Language	English
Publisher	The American Association of Immunologists
Publisher Date	2015-06-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Cell Migration Assays, Leukocyte Chemotaxis, Leukocyte Immunology Interleukin-8 Neutrophils Endothelium Cytology Glycosaminoglycans Metabolism Inflammation Discipline Immunology
Content Type	Text
Resource Type	Article
Subject	Immunology and Allergy Immunology

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